Detection of Human/Mouse/Rat MKK3 by Western Blot.
Western blot shows lysates of MDA-MB-468 human breast cancer cell line, TS1 mouse helper T cell line, and NRK rat normal kidney cell. PVDF membrane was probed with 1 µg/mL Mouse Anti-Human/Mouse/Rat MKK3 Monoclonal Antibody (Catalog # MAB2515) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, recombinant human MKK3 and MKK6 (2 ng/lane) were included. A specific band for MKK3 was detected at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
MKK3 in HeLa Human Cell Line.
MKK3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat MKK3 Monoclonal Antibody (Catalog # MAB2515) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C, as supplied.
1 month, 2 to 8 °C under sterile conditions after opening.
6 months, -20 to -70 °C under sterile conditions after opening.
Mitogen-activated protein kinase kinase 3 (MKK3), also known as MEK3, and the closely related MKK6 are activated by proinflammatory cytokines and environmental stresses. Activation occurs through phosphorylation by several upstream MAPK kinase kinases, including ASK1 and TAK1. Both MKK3 and MKK6 are dual specificity protein kinases, phosphorylating and activating the p38 MAP kinases at Thr and Tyr positions within the phosphoacceptor sequence Thr-Gly-Tyr.
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